Process Proce ss Bioch Biochemist emistry ry 46 (2011) 1572–1 1572–1578 578
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Process Biochemistry j o u r n a l h o m e p a g e : w w w . e l s e v i e r . c o m / l o c a t e / p r o c b i o
Biosynthesis and characterization of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) and poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) copolymers using using jatropha jatropha oil as the main carbon source Ko-Sin Ng a , Yoke-Ming Wong a , Takeharu Tsuge b , Kumar Sudesh a,∗ a b
Ecobiomate Ecobiomaterial rial Research Research Laboratory Laboratory,, School School of Biologica Biologicall Science, Science, Universiti Universiti Sains Sains Malaysia, Malaysia, 11800 Minden, Minden, Penang, Penang, Malaysia Malaysia Department Department of Innovative Innovative and Engineered Engineered Materials, Materials, Tokyo Institute Institute of Technology Technology,, 4259 Nagatsuta, Nagatsuta, Midori-ku, Midori-ku, Yokohama Yokohama 226-8502, 226-8502, Japan
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Article history: Receiv Received ed 8 Decemb December er 2010 2010 Receiv Received ed in revise revised d form form 4 April April 2011 2011 Accept Accepted ed 26 April April 2011 2011 Keywords: Jatropha oil Physic Physic nut Jatropha curcas Poly(3-hydroxybutyrate-co-3hydroxyvalerate) Poly(3-hydroxybutyrate-co-3hydroxyhexanoate) Bioplastics
a b s t r a c t
Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] and poly(3-hydroxybutyrate-co-3hydroxyhexanoate) [P(3HB-co-3HHx)] copolymers were produced using jatropha using jatropha oilas the carbon source. P(3HB-co-3HV) with a 3HV 3HV monomer as high as 42 mol% was produced by wild-type Cupriavidus necator H16 from a mixtur mixture e of jatropha jatropha oil oil and sodium valerate, and P(3HB-co-3HHx) with a 3HHx monomer of necator r PHB− 4/pBBREE32d13 harboring Aeromonas caviae PHA 3 mol% mol% was produced by transformant C. necato synthase, using jatropha oil oil as the sole carbon source. The results of differential scanning calorimetry, thermogravimetric analysis and gel permeation chromatography revealed that the copolymers produced from from jatroph jatropha a oil oil were were essentially the same as thos those e produced from other, more established carbon sources, such as sugars and other plant oils. This study demonstrates that jatropha oil oil is a potent potential ial necator and its transformant. renewable carbon source for the large-scale production of copolymers of copolymers by C. necator transformant.
1. Intr Introd oduc ucti tion on
Polyhy Polyhydro droxya xyalka lkano noate ate (PHA) (PHA) is a polyme polymerr that that is accumu accumulat lated ed by bact bacter eria ia as a carb carbon on and and ener energy gy stor storag age e mate materi rial al in the the pres pres-ence ence of an excess excess amoun amountt of carbo carbon n and and a limite limited d supplyof supplyof anothe anotherr nutrie nutrient nt,, which which can be in the form form of nitrog nitrogen, en, sulfur sulfur,, phosp phospho horus rus,, magn magnes esiu ium m or oxyg oxygen en  .. Among Among variou variouss known known biodeg biodegrad radabl able e poly olymeric eric mater ateria ials ls,, PHA PHA is know known n to be a ful fully biod iodegra egrada dabl ble e alternati alternative ve to conventi conventional onal plastics plastics  .. In terms terms of its applic applicati ations ons,, PHAis a good good materi material al for biopla bioplasti stics cs and and implan implantt biomat biomateri erialsdue alsdue to its biodegrad biodegradabili ability, ty, biocompa biocompatibil tibility, ity, thermopr thermoprocess ocessibili ibility ty and sustainability  .. Althou Although gh PHA is a good good substi substitut tute e for petrol petroleum eum-ba -based sed plasti plastics, cs, the the high high cost cost of prod produc ucti tion on make makess PHA PHA more more expe expens nsiv ive e than than conconventiona ventionall plastics plastics  .. In an effo effort rt to redu reduce ce the the prod produc ucti tion on cost cost of PHAs PHAs,, vari variou ouss carb carbon on sour source ces, s, such such as plan plantt oils oils and and suga sugars rs,, have have been been explor explored ed foruse in PHAproduct PHAproduction ion.. To date, date, theuse of variou variouss plant plant oils, oils, includ including ing soybea soybean n oil  ,, palm palm oil oil [6,7] [6,7],, coconu coconutt oil  and and corn corn oil oil  ,, has has been been repo report rted ed to resu result lt in high high yiel yields ds of PHA. PHA.
Corres Correspon pondin ding g author author.. Tel.: Tel.: +60 4 653436 6534367; 7; fax: fax: +60 4 6565125 6565125.. E-mail E-mail address: address: [email protected]
(K. Sudesh). Sudesh).
1359-5 1359-5113 113/$ /$ – seefront matter© matter© 2011ElsevierLtd. 2011ElsevierLtd. All rightsreser rightsreserved ved.. doi:10.1016/j.procbio.2011.04.012 doi:10.1016/j.procbio.2011.04.012
© 2011 Elsevier Ltd. All rights reserved.
Howe Howeve ver, r, the the use use of thes these e edib edible le oils oils as prim primar ary y carb carbon on sour source cess for biopla bioplasti sticc produc productio tion n may not not be sust sustai aina nabl ble e  .. Continuous conv conver ersi sion on of edib edible le oils oils to biop biopla last stic icss may may lea lead to a shor shorta tag ge in food food suppli supplies es and cause cause inﬂati inﬂation on in certai certain n count countrie ries, s, especi especiall ally y in third third wor world ld count countrie ries. s. Theref Therefore ore,, ourstudyaims to evalua evaluate te theuse of jatr jatrop opha ha oil, oil, whic which h is a nonnon-ed edib ible le plan plantt oil, oil, for for the the prod produc ucti tion on of PHA PHA copo copoly lyme mers rs.. In a prev previo ious us stud study, y, we showed showed that that jatrop jatropha ha oil oil can can be used used by Cupriavid grow and synthe synthesiz size e Cupriavidus us necator necator H16 to grow poly(3-hydroxybutyrate) poly(3-hydroxybutyrate) [P(3HB)] homopolymer  .. However, However, it is notknown notknown if jatrop jatropha ha oilaffects oilaffects the the incorp incorpora oratio tion n of comono comonomer mer and the the proper propertie tiess of PHA copoly copolymer mers. s. Jatropha curcas, also known known as physi physicc nut,is nut,is a multip multipurp urposeshru oseshrub b that that canreach canreach a heightof heightof 20ft and has glabro glabrous us branc branchle hlets ts  .. The life life expect expectanc ancy y of J. J. curcas is almost50 almost50 years,and years,and this this shrubis shrubis distri distribut butednativ ednativelyin elyin Centra Centrall and Sout South h Amer Americ ica, a, incl includ udin ing g in Mexi Mexico co,, Braz Brazil il and and Arge Argent ntin ina a and and can can now now also also be foun found d in many many part partss of Afri Africa ca and and Asia Asia  .. J . curca curcass is charac character terize ized d as a hardy hardy,, highly highly adapta adaptable ble,, drough drought-r t-resi esista stant nt crop crop and consequen consequently tly has high ecologic ecological al adaptabi adaptability. lity. Furthermor Furthermore, e, J. curcas is a dise diseas asee-re resi sist stan antt plan plantt beca becaus use e only only a few few inse insect ctss or fungi fungi cantransmit cantransmit their their diseas diseases es to the plant plantss  .. Jatroph atropha a oil can be consid considere ered d to be a biodie biodieselfuel selfuel becaus because e it has has a lowoil visco viscosit sity y whencompared whencompared to soybean,cotton soybean,cottonseedand seedand sunﬂoweroils sunﬂoweroils  .. The blackish J. curcas seeds seeds contai contain n ﬁve main main toxin toxins, s, whichare whichare phorbo phorboll
K.-S. Ng et al. / Process Biochemistry46 (2011) 1572–1578
Table 1 Biosynthesis of P(3HB- co-3HV) copolymer by C. necator H16 using the mixture of jatropha oil as the main carbon source and sodium valerate or sodium propionate as the precursor carbon sources.a Data shown are the means of triplicate. Mean data accompanied by different superscript alphabets are signiﬁcantly different (Tukey’s HSD test, p